Oxfendazole Nitazoxanide combination in experimental neurocysticercosis - Anti-inflammatory and cysticidal effects.

Neurocysticercosis (NCC) is a parasitic infection caused by the larval stage of the pork tapeworm, Taenia solium. The complications of NCC include seizures, headaches, cognitive impairment, and focal neurological deficits. In addition to antiparasitic drugs and surgery, the management of NCC includes the use of corticosteroids to reduce inflammation and control symptoms. The traditional treatment with albendazole and praziquantel has not been altered over 30 years and present several side effects. There are other anti-helminthic drugs such as oxfendazole and nitazoxanide that may show efficacy in NCC treatment. The aim of this study was to determine the histopathologic aspects of experimental NCC after in vivo treatment with the combination of oxfendazole and nitazoxanide. Balb/c mice were infected with T. crassiceps cysticerci and divided into groups of 10 animals each that received a single dose through gavage as follows: group treated with NaCl 0.9% (control group); group treated by monotherapy of the anti-helminthic drugs, 30 mg/kg in single dose of oxfendazole (OXF) or nitazoxanide (NTZ); and groups treated with the combination of the drugs (OXF/NTZ group). Macroscopic and microscopic analysis were performed. There was greater presence of final stage cysticerci after treatment. The microscopic analysis of the general pathological processes showed that the monotherapy with all treatment groups induced higher perivasculitis than what was observed in the control group. In contrast, the combination treatment showed a lower observation of PMN and MN inflammatory infiltration in comparison to the other treatments and to the control one. These results show that indeed the association of benzimidazole derivatives which present both anti-helminthic and anti-inflammatory properties with other cysticidal drugs are beneficial for the NCC treatment in which the aim is to destroy parasite without inducing inflammatory damage in the brain tissue.

Oxfendazole induces protein catabolism and gluconeogenesis in experimental neurocysticercosis.

Neurocysticercosis (NCC) is an endemic public health disease of the central nervous system highly related to epilepsy and seizures. Taenia crassiceps is an experimental model used for NCC and biochemical studies of the host-parasite relationship. For the past 50 years the NCC therapeutic treatment is performed with albendazole (ABZ) and praziquantel which opens a gap for new therapies due to parasitic resistance and other adverse effects of the drugs. Oxfendazole (OXF) is an albendazole derivative with efficacy against tissue cestodes of veterinary importance. The aim of this study was to determine the metabolic impact of OXF on T. crassiceps cysticerci intracranially inoculated in Balb/C mice. The animals were intracranially inoculated with T. crassiceps cysticerci and 30 days later received single dose oral treatment of OXF, ABZ and NaCl 0.9% (control group). The metabolic impact was quantified through the detection of metabolites from glycolysis, anaerobic fermentation of lactate and propionate, tricarboxylic acid cycle, protein catabolism, fatty acids oxidation. The differences observed in the concentrations of metabolites from the OXF treated group showed that the drug induced gluconeogenesis, increase in protein catabolism, fatty acids oxidation and propionate fermentation in comparison to the ABZ and control treated groups. In conclusion, OXF induced greater metabolic impact in T. crassiceps cysticerci than the standard NCC treatment, ABZ, showing that it may represent an alternative drug for its treatment.

IgG Avidity in Samples Collected on Filter Paper: Importance of The Early Diagnosis of Congenital Toxoplasmosis.

OBJECTIVE: The purpose of the present study is to standardize and evaluate the use of the immunoglobulin G (IgG) antibody avidity test on blood samples from newborns collected on filter paper to perform the heel test aiming at its implementation in ongoing programs. METHODS: Blood samples from newborns were collected on filter paper simultaneously with the heel prick test. All samples were subjected to immunoglobulin M IgM and IgG enzyme-linked immunosorbent assays (ELISA). Peripheral blood was collected again in the traditional way and on filter paper from newborns with high IgG levels (33). Three types of techniques were performed, the standard for measuring IgG in serum, adapted for filter paper and the technique of IgG avidity in serum and on filter paper. The results of the avidity test were classified according to the Rahbari protocol. RESULTS: Among the 177 samples, 17 were collected in duplicate from the same child, 1 of peripheral blood and 1 on filter paper. In this analysis, 1 (5.88%) of the 17 samples collected in duplicate also exhibited low IgG avidity, suggesting congenital infection. In addition, the results obtained from serum and filter paper were in agreement, that is, 16 (94.12%) samples presented high avidity, with 100% agreement between the results obtained from serum and from filter paper. CONCLUSION: The results of the present study indicate that the avidity test may be another valuable method for the diagnosis of congenital toxoplasmosis in newborns.

OBJETIVO: O objetivo do presente estudo é padronizar e avaliar a utilização do teste de avidez de anticorpos imunoglobulina G (IgG) em amostras de sangue de recém-nascidos (RNs) coletadas em papel filtro para a realização do teste do pezinho visando a implementação nos programas já vigentes. MéTODOS: Foram coletadas amostras de sangue de recém-nascidos em papel filtro simultaneamente ao teste do pezinho. Em todas as amostras, foram realizados os testes imunoenzimáticos (ELISA) imunoglobulina M (IgM) e IgG. Dos RNs que apresentaram altos índices de IgG (33), foi novamente coletado sangue periférico da forma tradicional e em papel filtro. Foram realizadas técnicas padrão para a dosagem de IgG em soro, adaptadas para papel filtro, e a técnica de avidez de IgG em soro e em papel filtro. Os valores obtidos para o teste de avidez foram classificados de acordo com o protocolo de Rahbari. RESULTADOS: Dentre as 177 recoletas, em 17 amostras foi realizada a coleta simultânea de sangue periférico e papel filtro da mesma criança. Nesta análise, 1 (5,88%) das 17 amostras coletadas em duplicata obteve também baixa avidez de IgG, sugerindo infecção congênita da criança, e houve concordância entre os resultados obtidos em soro e em papel filtro: 16 (94,12%) das amostras apresentaram alta avidez, com concordância de 100% entre os resultados obtidos em soro e em papel filtro. CONCLUSãO: Os dados do presente trabalho evidenciam que o teste de avidez poderá ser mais um método valioso a ser utilizado no diagnóstico da toxoplasmose congênita em RNs.

IgG Avidity in Samples Collected on Filter Paper: Importance of The Early Diagnosis of Congenital Toxoplasmosis / Avidez de IgG em amostras coletadas em papel filtro: Importância no diagnóstico precoce da toxoplasmose congênita

Abstract Objective The purpose of the present study is to standardize and evaluate the use of the immunoglobulin G (IgG) antibody avidity test on blood samples from newborns collected on filter paper to perform the heel test aiming at its implementation in ongoing programs. Methods Blood samples from newborns were collected on filter paper simultaneously with the heel prick test. All samples were subjected to immunoglobulin M IgM and IgG enzyme-linked immunosorbent assays (ELISA). Peripheral blood was collected again in the traditional way and on filter paper from newborns with high IgG levels (33). Three types of techniques were performed, the standard for measuring IgG in serum, adapted for filter paper and the technique of IgG avidity in serum and on filter paper. The results of the avidity test were classified according to the Rahbari protocol. Results Among the 177 samples, 17 were collected in duplicate from the same child, 1 of peripheral blood and 1 on filter paper. In this analysis, 1 (5.88%) of the 17 samples collected in duplicate also exhibited low IgG avidity, suggesting congenital infection. In addition, the results obtained from serum and filter paper were in agreement, that is, 16 (94.12%) samples presented high avidity, with 100% agreement between the results obtained from serum and from filter paper. Conclusion The results of the present study indicate that the avidity test may be another valuable method for the diagnosis of congenital toxoplasmosis in newborns.

Resumo Objetivo O objetivo do presente estudo é padronizar e avaliar a utilização do teste de avidez de anticorpos imunoglobulina G (IgG) em amostras de sangue de recémnascidos (RNs) coletadas em papel filtro para a realização do teste do pezinho visando a implementação nos programas já vigentes. Métodos Foram coletadas amostras de sangue de recém-nascidos em papel filtro simultaneamente ao teste do pezinho. Em todas as amostras, foram realizados os testes imunoenzimáticos (ELISA) imunoglobulina M (IgM) e IgG. Dos RNs que apresentaram altos índices de IgG (33), foi novamente coletado sangue periférico da forma tradicional e em papel filtro. Foram realizadas técnicas padrão para a dosagem de IgG em soro, adaptadas para papel filtro, e a técnica de avidez de IgG em soro e em papel filtro. Os valores obtidos para o teste de avidez foram classificados de acordo com o protocolo de Rahbari. Resultados Dentre as 177 recoletas, em 17 amostras foi realizada a coleta simultânea de sangue periférico e papel filtro da mesma criança. Nesta análise, 1 (5,88%) das 17 amostras coletadas em duplicata obteve também baixa avidez de IgG, sugerindo infecção congênita da criança, e houve concordância entre os resultados obtidos em soro e em papel filtro: 16 (94,12%) das amostras apresentaram alta avidez, com concordância de 100% entre os resultados obtidos em soro e em papel filtro. Conclusão Os dados do presente trabalho evidenciam que o teste de avidez poderá ser mais um método valioso a ser utilizado no diagnóstico da toxoplasmose congênita em RNs.

Molecular characterization of Toxoplasma gondii isolates from free-range chickens reveals new genotypes in Goiânia, Goiás, Brazil.

The aim of this study was to evaluate the genotypic characteristics of Toxoplasma gondii isolated from free-range chickens in the metropolitan area of Goiânia, Goiás, in Brazil's central-west region. The seroprevalence rate was found to be 96%, according to an indirect hemagglutination assay. Brain and heart samples were processed by peptic digestion for a mice bioassay. The tissues were homogenized and the resulting samples were subjected to polymerase chain reaction (PCR), which revealed that 64% of them contained the parasite's DNA. The mice bioassay revealed 15 isolates, 8 of them tachyzoites isolates from the peritoneal lavage and 7 from brain cysts. T. gondii genotypes were determined through PCR-RFLP, using the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico and CS3. Three genotypes were identified, inclued ToxoDB #65, and the other two are not yet described in the literature. Hence, we conclude that the isolates obtained from the metropolitan area of Goiânia showed relatively low genetic diversity.

Identification of Biomarkers for Diagnosis and Prognosis of Congenital and Acute Toxoplasmosis.

BACKGROUND: The diagnosis of congenital toxoplasmosis can be inconclusive in many cases. Despite the several serological tests developed, the literature on biomarkers that can assist in the diagnosis of congenital an acute toxoplasmosis is limited. The objective of this study was to analyze the immunoreactive profile of Toxoplasma gondii protein bands with the potential to be biomarkers for diagnosis and prognosis of congenital and acute toxoplasmosis. METHODS: Peripheral blood samples from women of childbearing age and/or pregnant women diagnosed with acquired toxoplasmosis as well as from congenitally infected children were selected and submitted to immunoblotting for analysis of the immunoreactive bands profile by immunoglobulin G (IgG) antibodies. RESULTS: When comparing the immunoreactive bands profile for antibodies present in samples from different groups and subgroups, the 150, 18.5, and 16.96-kDa bands were more immunoreactive with the antibodies present in serum samples from the acquired infection group. The 343, 189, 150, 75, and 42-kDa bands showed more chance to be detected by the symptomatic congenital infection subgroup samples, while the 61, 50, and 16.96-kDa bands were significantly immunoreactive with the acute infection subgroup samples. CONCLUSIONS: The identification of these potential biomarkers can assist in early diagnosis and treatment of congenital toxoplasmosis.

Partial inhibition of the main energetic pathways and its metabolic consequences after in vivo treatment with benzimidazole derivatives in experimental neurocysticercosis.

Benzimidazole derivatives such as albendazole (ABZ) and mebendazole are important molecules used in helminthic treatment. Neurocysticercosis is the main cause of acquired epilepsy throughout the world and is currently treated with ABZ. New molecules have been studied in order to aid in the treatment of this neglected tropical disease, among them RCB15 and RCB20. The aim of this study was to evaluate the metabolic impact of RCB15 and RCB20 on Taenia crassiceps cysticerci intracranially inoculated in Balb/c mice. Thirty days after the inoculation the mice were treated with 50 mg kg-1 of RCB15, RCB20, ABZ or NaCl 0.9%. The euthanasia and cysticerci removal were performed 24 h after the treatment. The cysticerci were analysed through high performance liquid chromatography. After the treatments, there was an impairment in the main energetic pathways such as glycolytic pathway, homolactic fermentation or in mitochondrion energy production detected through the decrease in pyruvate, lactate, oxaloacetate, malate and fumarate concentrations. This induced the parasite to resort to alternative energetic pathways such as proteins catabolism, propionate fermentation and fatty acids oxidation. Therefore, benzimidazole derivatives are a promising alternative to ABZ use as they also reach the brain tissue and induce a metabolic stress in the cysticerci.

Basic heel prick test: inclusion of screening, diagnosis and criteria for early confirmation of congenital infection by Toxoplasma gondii.

Toxoplasma gondii can cross the placental barrier, causing fetal infection with potentially severe sequelae. The aim of this study was to evaluate whether the serological screening for toxoplasmosis should be included in the basic neonatal heel prick test in order to establish criteria for the confirmation and/or exclusion of the diagnosis of congenital infection in newborns treated at three public health units in the metropolitan region of Goiania, Goias State, Brazil. Blood samples were collected on filter paper from newborns and later, peripheral blood samples from the mothers and their respective children were obtained to confirm or exclude the diagnosis of suspected congenital infection, by means of an enzyme-linked immunosorbent assay (IgM and IgG) and a polymerase chain reaction assay. From a total of 1,159 blood samples collected on filter paper, 43.92% were reactive to IgG and 0.17% to anti-T. gondii IgM and IgG. One hundred and twenty-seven paired samples (mother and child) were collected following consensual protocols for peripheral blood collection. Results obtained from the filter paper and peripheral blood of the newborns were 90.55% concordant. A comparison of the mother and child blood test results showed agreement regarding the detection of IgG in 90.48% of the samples. The parasite DNA was detected in the peripheral blood of one child. In view of the results obtained in this study, the inclusion of the serological screening for toxoplasmosis in the newborn heel prick test proved to be effective for the early detection of congenital T. gondii infection.

Nursing infant with acquired toxoplasmosis in the first months of life - a case report.

Toxoplasmosis is caused by Toxoplasma gondii and the probability of this infection occurring in the first months of life is usually low because its transmission is related to eating habits. A 6-month-old nursing infant was diagnosed with acute toxoplasmosis, which was identified through anti- T. gondii IgA, IgM and low-avidity IgG serologic assays, polymerase chain reaction (PCR) and mouse bioassay test although its mother was seronegative. This serological divergence between mother and child led us to interview the mother regarding epidemiological factors. During this interview, she reported that she had given her 2-month-old baby a piece of undercooked beef to suck on. After some time, the baby presented fever and cervical lymphadenitis. This report emphasizes the importance of serological surveys of toxoplasmosis in nursing infants presenting with fever and lymphadenitis, in view of the possible acquisition of toxoplasmosis in the first months of life.